Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, Washington 99164, USA.
Foodborne Pathog Dis. 2013 Jul;10(7):624-31. doi: 10.1089/fpd.2012.1313. Epub 2013 Jun 6.
Campylobacter jejuni is one of the most important causes of foodborne gastroenteritis. Chickens are considered a reservoir host of C. jejuni, and epidemiological studies have shown that contaminated chicken meat is a primary source of human infection. The objective of this study was to produce chicken egg-yolk-derived antibody (IgY) against the five C. jejuni colonization-associated proteins or CAPs (CadF, FlaA, MOMP, FlpA, and CmeC). Recombinant C. jejuni CAPs were expressed in Escherichia coli and were purified by affinity chromatography. Specific-pathogen-free laying hens were hyperimmunized with each recombinant CAP to induce production of α-CAP-specific IgY. Egg yolks were collected from immunized and nonimmunized hens and were lyophilized to obtain egg-yolk powder (EYP) with or without α-C. jejuni CAP-specific IgY. IgY was purified from EYP, and the antibody response in serum and egg yolk was tested by indirect enzyme-linked immunosorbent assay. The α-C. jejuni CAP-specific IgY levels were significantly (p<0.05) higher in both serum and EYP obtained from immunized hens as compared with the nonimmunized hens. Each α-C. jejuni CAP-specific IgY reacted with the C. jejuni cells and recombinant CAPs as detected by immunofluorescence microscopy and Western blot assays, respectively. We also show that α-CadF, α-MOMP, and α-CmeC IgY significantly reduced adherence of C. jejuni to the chicken hepatocellular carcinoma (LMH) cells, suggesting that these α-C. jejuni CAP-specific IgY may be useful as a passive immunotherapeutic to reduce C. jejuni colonization in chickens.
空肠弯曲菌是食源性肠胃炎最重要的病因之一。鸡被认为是空肠弯曲菌的储存宿主，流行病学研究表明，受污染的鸡肉是人感染的主要来源。本研究的目的是生产针对 5 种空肠弯曲菌定植相关蛋白（CadF、FlaA、MOMP、FlpA 和 CmeC）的鸡卵黄源性抗体（IgY）。重组空肠弯曲菌 CAP 用大肠杆菌表达并通过亲和层析纯化。将无特定病原体的产蛋母鸡用每种重组 CAP 进行超免疫，以诱导产生针对 α-CAP 的特异性 IgY。从免疫和非免疫母鸡收集卵黄并冻干，以获得含有或不含有 α-C. jejuni CAP 特异性 IgY 的卵黄粉（EYP）。从 EYP 中纯化 IgY，并通过间接酶联免疫吸附试验检测血清和卵黄中的抗体反应。与非免疫母鸡相比，免疫母鸡的血清和 EYP 中的 α-C. jejuni CAP 特异性 IgY 水平均显著升高（p<0.05）。每个 α-C. jejuni CAP 特异性 IgY 通过免疫荧光显微镜和 Western blot 检测分别与空肠弯曲菌细胞和重组 CAP 反应。我们还表明，α-CadF、α-MOMP 和 α-CmeC IgY 显著降低了空肠弯曲菌对鸡肝癌（LMH）细胞的黏附，表明这些 α-C. jejuni CAP 特异性 IgY 可用作被动免疫疗法来减少鸡中的空肠弯曲菌定植。
